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目的 :检测TFP对培养的豚鼠RPE细胞cAMP浓度和PKA活性的影响。探讨CaM被抑制时 ,与cAMP -PKA信号系统相关性及在RPE细胞增殖调控中的作用。从信号传导角度探讨TFP抑制RPE细胞迁移和增殖的机制。方法 :1 应用12 5I -cAMP试剂盒 ,采用放免法测定 10umol/LTFP作用不同时间RPE细胞内cAMP浓度的变化。 2 应用液闪法测定 10umol/LTFP作用不同时间后 ,RPE细胞PKA活性的变化。结果 :1 TFP对豚鼠RPE细胞cAMP浓度的影响 :统计结果表明 ,与对照组cAMP值 ( 0 410 9± 0 0 931pmol/5 0ul)相比 ,TFP作用 5分钟内即引起cAMP快速的升高达 ( 0 80 5 1± 0 5 5 3pmol/5 0ul) (P <0 0 1) ,直到 3小时仍维持较高的水平 ( 0 72 85±0 0 2 94pmol/5 0ul) (P <0 0 5 )。 2 TFP对豚鼠RPE细胞PKA活性的影响 :统计结果表明 ,与对照组PKA活性值 ( 4 5 5 73±2 41cpm/mgprotiein)相比 ,TFP作用 5min内即引起PKA活性快速上升达 ( 135 6 5± 1 81cpm /mg protein) (P <0 0 1) ,直至3h仍维持较高水平 ( 6 5 95± 5 0 1cpm/mg protein) (P <0 0 5 )。结论 :1 在TFP抑制RPE细胞增殖过程中 ,cAMP -PKA信号系统呈上调趋势。 2 在RPE细胞增殖抑制过程中 ,Ca2 + -CaM系统与cAMP -PKA信号系统存在着拮抗关系。 3 在TFP引起cA
Objective: To detect the effect of TFP on cAMP concentration and PKA activity in cultured RPE cells of guinea pigs. To investigate the relationship between cAMP-PKA signaling system and the regulation of RPE cell proliferation when CaM is inhibited. To explore the mechanism of TFP in inhibiting RPE cell migration and proliferation from the perspective of signal transduction. Methods: 1 The changes of cAMP concentration in RPE cells were detected by radioimmunoassay using 125I-cAMP kit. The changes of PKA activity in RPE cells after 10umol / LTFP treatment were measured by liquid-flash method. Results: 1 The effect of TFP on cAMP concentration in RPE cells of guinea pigs: The statistical results showed that the cAMP rapidly increased within 5 minutes after TFP administration compared with the control group (0 410 9 ± 0 0 931 pmol / 5 0 ul) 0 80 5 1 ± 0 5 5 3 pmol / 5 0 ul) (P 0 01), up to 3 hours (0 72 85 ± 0 0 2 94 pmol 5 0 ul) (P 0 05) . The effect of TFP on the PKA activity of RPE cells in guinea pigs: The results showed that PKA activity rapidly increased within 5 min after TFP administration compared with PKA activity of control group (45 5 73 ± 2 41 cpm / mgprotiein) (135 6 5 ± 1 81 cpm / mg protein) (P <0.01), and remained high until 3 h (955 ± 5 0 1 cpm / mg protein) (P <0 05). Conclusions: 1 In the process of TFP inhibiting the proliferation of RPE cells, the cAMP-PKA signaling system is up-regulated. 2 There is an antagonistic relationship between Ca2 + -CaM system and cAMP-PKA signaling system in RPE cell proliferation. 3 causes cA at TFP